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First Mates Download: Enjoy the Best of Cecelia Dowdy's Christian Fiction



In the maritime field, it is vital to possess proper skills and abilities to succeed. Whether you are applying as a ship crew, deck officer, or captain, your resume needs to reflect those skills. This is especially important when applying for the first mate position. To help you with your job search tool, we have prepared some tips and a first mate resume example.




First Mates download



First mates in the US get a median salary of $51,334. That is around $25 per hour. It could vary depending on the company and credentials of the candidate. There has been a record of as high as $79,974 to as low as $26,088.


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Because little is known about non-mammalian Mates, our study was focused on the identification and initial characterization of Mates in zebrafish (Danio rerio), a well-established model species in biomedical and environmental research. Our primary goals were (1) to obtain a comprehensive set of phylogenetic and gene expression data to indicate the potential importance of Mates in zebrafish embryos and adults and (2) to determine the interaction profiles of zebrafish Mates with potential substrates as the basis for developing plausible hypotheses related to the physiological and/or defensive roles of these transporters. To position and annotate zebrafish Mates in relation to the other phyla, we performed a detailed phylogenetic analysis of MATEs/Mates from bacteria to mammals. After identification of six zebrafish mate genes, synteny analysis was performed in order to elucidate the orthologous relationships with human MATEs. Gene expression analysis (qRT-PCR) of the identified zebrafish mates was performed in zebrafish embryos at different developmental stages, followed by tissue-specific determinations in adults. Finally, an initial functional characterization of selected proteins was performed using transport activity assays with model substrates after heterologous overexpression of zebrafish Mates in HEK293T cells.


The embryonic and tissue-specific gene expression data for zebrafish mate genes are shown in Fig. 3A,B and additional graphical presentation of the tissue expression data are provided in the Supplementary material (Fig. S2). The expression levels of zebrafish Mates are described in the following paragraphs and were compared based on the expression thresholds for qRT-PCR data described in the Methods section. The expression of mate genes in zebrafish embryos in the early developmental stages showed differential expression of specific genes (Fig. 3A). At the first two time points (1 and 4 hpf, respectively), all genes showed low to moderate expression, probably indicating maternal transfer. With the onset of embryonic transcriptional activity at 6 hpf, a significant activation of two genes (mate6 and mate7) was observed, suggesting important roles in embryonic development. Between the finish of hatching at 72 hpf and the completion of organogenesis at 120 hpf, all genes except mate4 reached moderately high to high levels of expression (Fig. 3A).


In liver, mate6 and mate8 were expressed at moderately high expression levels, followed by moderate levels of mate7 in males. Mate7 showed notably higher expression in male liver (20 times). Mates generally showed lower expression in the gills than in other tissues. All genes were moderately expressed except for the low expression of mate6 and mate5 in females. Sex differences were observed for mate5 and mate7, which showed 10 and 2 times greater expression in males, respectively. In the intestine, the predominant mate genes are mate6 in males (high expression) and mate3 and mate5 in males (moderately high expression). In females, all genes were expressed at low to moderate levels. In the testes, mates were generally highly expressed: mate7 showed very high expression levels, followed by high expression of mate3, mate4, mate5 and mate8 and moderate expression of mate6. By contrast, in the ovaries, all genes showed low to moderate expression levels. In the zebrafish brain, mate7 and mate8 were highly expressed, as was mate6 in males, followed by moderately high expression of mate3 in males. Gender differences in mate expression in the brain were found for mate3 and mate6, which showed 6 times higher expression in males. In the eye, mates were generally highly expressed, with high expression levels of mate4 in females and mate7 in both genders, followed by moderately high expression of mate6 and mate8 in both genders and mate5 in females.


MATE/Mate transporters have recently been recognized as important components of the phase III cellular detoxification in humans, mice and rats7,15. However, knowledge of the non-mammalian Mate proteins was lacking. Therefore, we focused on elucidating their identity, gene expression and transport properties to obtain the first comprehensive data set to help determine the possible roles of these transporters in zebrafish, an important model vertebrate species.


We identified six mate genes in the zebrafish genome and found that fish Mate proteins clearly form a distinct cluster from tetrapod MATE/Mates (Fig. 1). Based on the phylogenetic analysis, direct orthologous relationships could not be determined among fish and mammalian MATE/Mate proteins. To address this obstacle, we performed synteny analysis which showed that zebrafish mates are syntenic to human MATE genes (Fig. 2). The observed differences in the surrounding gene environments are most likely a consequence of the teleost-specific whole genome duplication (WGD) that has been shown to form double conserved synteny blocks (DCS)34. DCS are defined35,36 as gene regions in non-duplicated species that are found on two different chromosomes in the species that underwent the WGD, although the genes may not be adjacent in the duplicated species. Therefore, considering the similar gene environments among the two zebrafish mate clusters, i.e., mate3, 4 and 6 on chromosome 21 and mate5, 7 and 8 on chromosome 15 (Fig. 2), we propose that these two zebrafish clusters arose from a genome duplication event.


All six zebrafish mates are ubiquitously expressed in the analyzed tissues of adult zebrafish. Unlike human, mouse and rat MATEs/mates, none of the six zebrafish genes predominates in a single tissue. Human MATE1 is present in several tissues and is the predominant MATE in kidney and liver. It is localized in the apical membrane of the proximal tubule cells and hepatocytes, where it mediates the efflux of compounds into urine and/or bile37. Mouse Mate1a and b isoforms have similar tissue distribution patterns with dominant presence in kidney. Mate1a is also present in the liver7,9,11, while rat Mate1 is found in the kidney and placenta4,12,38. Human MATE2 (and its variant MATE2-K) is predominantly expressed in the apical membrane of the proximal renal tubules, whereas mouse and rat Mate2 are predominantly found in the testes10 and have not been notably detected in other tissues37. In comparison, similar to mammalian MATEs/mates37 transcripts, the highest expression of zebrafish mates is found in the kidney and testes and to a lesser extent in the liver and brain (Fig. 3B). However, apart from these general similarities, the tissue expression pattern of each zebrafish gene cannot be directly matched to a human, mouse and/or rat MATE/mate transporter(s). Among zebrafish mates, mate6, 7 and 8 are the only ones present in both the kidney and liver, with moderate to very high expression levels in both genders, whereas mate4 and 5 are predominantly expressed in the testes and kidney and are not present in the liver at notable levels (Fig. 3B). In that sense, mate4 and mate5 resemble human MATE2, which is also not found in the liver and mouse and rat Mate2, which are found in the testes at high expression levels. Summarizing our tissue gene expression data and considering the high renal expression levels, we propose that all six zebrafish Mates are important transporters in fish kidney and Mate6 and Mate8 may be important in the liver, where they are found at moderate levels (Fig. 3B). In the testes, with the exception of mate6, all mates are highly expressed, indicating an important transport function in this tissue, possibly in testosterone transport as proposed previously10 for human MATEs.


In conclusion, this study reveals novel insights into the evolution of the MATE/Mate transporter family and provides the first comprehensive data set on Mate transporters in zebrafish, an important vertebrate model species. A sound methodological basis for future studies investigating the Mate interactors is presented. Finally, plausible hypotheses related to the possible roles of Mate proteins in the transport of physiological and/or xenobiotic substrates in zebrafish embryos and/or adults are proposed. They remain to be more specifically addressed in follow-up studies through detailed functional characterization of single transporter/s in suitable expression systems and zebrafish functional genomics studies using targeted gene knockdown.


How to cite this article: Lončar, J. et al. The first characterization of multidrug and toxin extrusion (MATE/SLC47) proteins in zebrafish (Danio rerio). Sci. Rep. 6, 28937; doi: 10.1038/srep28937 (2016).


Bowtie 2 is often thefirst step in pipelines for comparative genomics, including forvariation calling, ChIP-seq, RNA-seq, BS-seq. Bowtie 2 and Bowtie (also called "Bowtie 1" here) are also tightlyintegrated into many other tools, some of which arelisted here. 2ff7e9595c


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